Project 4: Application of Comparative Genomics, Transcriptomics, & Proteomics Opt

这项提议将应用先进的分子工具来理解和优化微生物 常见的超级基金污染物全氯乙烯(PCE)和三氯乙烯(TCE)的解毒 对人类和生态健康构成重大威胁。通过研究基本过程 负责PCE和TCE的厌氧微生物降解，这项工作将促进改良现场 生物修复过程。之前由NIEHS资助的研究试图识别和理解 复杂群落中负责脱氯的微生物。这项工作采取了下一个合乎逻辑的步骤 通过专注于唯一的细菌属，脱卤球菌属，已知可以完全将PCE和TCE降低到 乙烯。将使用全基因组微阵列、蛋白质组分析和定量聚合酶链式反应来表征 嗜盐球菌属不同菌株的基因组差异及基因表达评价 以及由各种底物的还原脱氯引起的蛋白质组变化，在简单和 复杂的微生物群落，以及其他生理扰动。基因组和转录组 不同降解能力的脱卤球菌菌株的比较将确定途径 负责特定和一般的新陈代谢，以及揭示 各种分离的菌株。脱氢球菌菌株在纯培养和混合培养中的转录比较 将确定参与物种间相互作用的途径，揭示营养需求和代谢作用 关于财团中的脱卤球菌属，并解决生物修复应用方面的限制 分离的脱卤球菌菌株生长不良。来自菌株鉴定、基因表达和 蛋白质生产将符合动力学模型，可以用来预测还原速度 特征不佳的微生物群落的脱氯作用。这项研究将达到SBRP的目标 限制在超级基金地点常见的化学物质的人体暴露和毒性，方法是促进 PCE和TCE原位生物修复技术的研究进展 无需提取到地表的地下位置，避免了潜在的人类和生态暴露。

This proposal will apply advanced molecular tools to understand and optimize the microbial detoxification of common Superfund pollutants, perchloroethene (PCE) and trichloroethene (TCE), which pose a significant threat to human and ecological health. By studying the fundamental processes responsible for anaerobic microbial degradation of PCE and TCE, this work will promote improved situ bioremediation processes. Previous NIEHS-funded research sought to identify and understand the microbes within complex communities responsible for dechlorination. This work takes the next logical step by focusing on the only genus of bacteria, Dehalococcoides, known to completely reduce PCE and TCE to ethene. Whole-genome microarrays, proteomic analyses, and quantitative PCR will be used to characterize the genomic differences between a variety of Dehalococcoides strains and to evaluate gene expression and proteomic changes caused by reductive dechlorination of a variety of substrates, growth in simple and complex microbial communities, and other physiological perturbations. Genomic and transcriptomic comparison of Dehalococcoides strains with different degradation abilities will identify the pathways responsible for specific and general metabolism as well as reveal the evolutionary relationship between the various isolated strains. Transcriptomic comparison of Dehalococcoides strains in pure and mixed cultures will identify pathways involved in inter-species interactions, reveal the nutritional needs and metabolic roles of Dehalococcoides in consortia, and address the limitation in bioremediation applications presented by the poor growth of isolated Dehalococcoides strains. Data from strain identification, gene.expression, and protein production will be complied into kinetic models that can be used to predict rates of reductive dechlorination by poorly characterized microbial communities. This research will meet the SBRP goal of limiting the human exposure and toxicity of chemicals commonly found at Superfund sites by advancing the development of in situ bioremediation of PCE and TCE, a technology that destroys contaminants in their subsurface location without extraction to the surface, avoiding potential human and ecological exposure.